Endbulbs Of Held Research Articles

Frequency-Independent Synaptic Transmission Supports a Linear Vestibular Behavior

The vestibular system is responsible for transforming head motion into precise eye, head, and body movements that rapidly stabilize gaze and posture. How do central excitatory synapses mediate behavioral outputs accurately matched to sensory inputs over a wide dynamic range? Here we demonstrate that vestibular afferent synapses in vitro express frequency-independent transmission that spans their in vivo dynamic range (5-150 spikes/s). As a result, the synaptic charge transfer per unit time is linearly related to vestibular afferent activity in both projection and intrinsic neurons of the vestibular nuclei. Neither postsynaptic glutamate receptor desensitization nor saturation affect the relative amplitude or frequency-independence of steady-state transmission. Finally, we show that vestibular nucleus neurons can transduce synaptic inputs into linear changes in firing rate output without relying on one-to-one calyceal transmission. These data provide a physiological basis for the remarkable linearity of vestibular reflexes.

Temporal Coding by Cochlear Nucleus Bushy Cells in DBA/2J Mice with Early Onset Hearing Loss

The bushy cells of the anterior ventral cochlear nucleus (AVCN) preserve or improve the temporal coding of sound information arriving from auditory nerve fibers (ANF). The critical cellular mechanisms entailed in this process include the specialized nerve terminals, the endbulbs of Held, and the membrane conductance configuration of the bushy cell. In one strain of mice (DBA/2J), an early-onset hearing loss can cause a reduction in neurotransmitter release probability, and a smaller and slower spontaneous miniature excitatory postsynaptic current (EPSC) at the endbulb synapse. In the present study, by using a brain slice preparation, we tested the hypothesis that these changes in synaptic transmission would degrade the transmission of timing information from the ANF to the AVCN bushy neuron. We show that the electrical excitability of bushy cells in hearing-impaired old DBA mice was different from that in young, normal-hearing DBA mice. We found an increase in the action potential (AP) firing threshold with current injection; a larger AP afterhyperpolarization; and an increase in the number of spikes produced by large depolarizing currents. We also tested the temporal precision of bushy cell responses to high-frequency stimulation of the ANF. The standard deviation of spikes (spike jitter) produced by ANF-evoked excitatory postsynaptic potentials (EPSPs) was largely unaffected in old DBA mice. However, spike entrainment during a 100-Hz volley of EPSPs was significantly reduced. This was not a limitation of the ability of bushy cells to fire APs at this stimulus frequency, because entrainment to trains of current pulses was unaffected. Moreover, the decrease in entrainment is not attributable to increased synaptic depression. Surprisingly, the spike latency was 0.46 ms shorter in old DBA mice, and was apparently attributable to a faster conduction velocity, since the evoked excitatory postsynaptic current (EPSC) latency was shorter in old DBA mice as well. We also tested the contribution of the low-voltage-activated K+ conductance (g (KLV)) on the spike latency by using dynamic clamp. Alteration in g (KLV) had little effect on the spike latency. To test whether these changes in DBA mice were simply a result of continued postnatal maturation, we repeated the experiments in CBA mice, a strain that shows normal hearing thresholds through this age range. CBA mice exhibited no reduction in entrainment or increased spike jitter with age. We conclude that the ability of AVCN bushy neurons to reliably follow ANF EPSPs is compromised in a frequency-dependent fashion in hearing-impaired mice. This effect can be best explained by an increase in spike threshold.

Development of synapses and expression of a voltage-gated potassium channel in chick embryonic auditory nuclei

The potassium channel protein, Kv3.1, is abundantly expressed in the chick auditory pathway. Its b-isoform is found in nucleus magnocellularis, which receives the cochlear input, both before and after the establishment of synaptic connections. It is also present in cell cultures in the absence of any peripheral input. However, the expression of this isoform in the embryo has been shown to increase with development. Here, we address the question of the correlation between maturation of synapses in the auditory pathway and the pattern of expression of the b-isoform in a series of embryos prepared for immunohistochemistry at Hamburger–Hamilton stages equivalent to E10, E12, E14, and E17. We show here that this subunit translocates from the perinuclear cytoplasm to the cell membrane domain in nucleus magnocellularis at the time that cochlear nerve endings emerge as endbulbs of Held (E17). In nucleus laminaris, by this time, while abundant Kv3.1b occurs in the perinuclear cytoplasm, a translocation to the cell membrane domain has not yet occurred, and the mature peri-synaptic localization is delayed to a later stage. This difference suggests a hierarchy in the developmental expression of Kv3.1. An unexpected finding is the expression of the a-isoform of Kv3.1 in astrocytes, especially those which surround the developing nuclei and their connecting fibers. We also report here for the first time the presence of Kv3.1b in the initial segments of axons at the times when they begin to form. Our observations suggest that the Kv3.1 channel protein is regulated through mechanisms linked to the development of synaptic activity.

Postnatal development of a large auditory nerve terminal: The endbulb of Held in cats

The endbulbs of Held are formed by the ascending branches of myelinated auditory nerve fibers and represent one of the largest synaptic endings in the brain. Most of the developmental changes in structure occur during the first 30 postnatal days of age. The neonatal endbulb begins as a flattened expansion with many filopodia, resembling a growth cone and characterized by numerous puncta adherentia and synapses associated with small postsynaptic densities; the most impressive feature of the ending at this age is its highly irregular plasma membrane that interdigitates with that of the postsynaptic spherical bushy cell. During these first 30 days, the number of puncta adherentia diminishes, postsynaptic densities nearly double in size, intermembraneous cisternae emerge, and plasma membranes flatten. These features endow the endbulb with an adult-like appearance. On the other hand, synaptic vesicle density increases progressively from approximately 50/μm 2 at birth to 100/μm 2 at adulthood. Mitochondria size remains constant over this developmental period but mitochondrial volume fraction increases until 60 days postnatal. Although many features of endbulb morphology stabilize by 30 days, other features suggest that endbulb development continues into the third month of age. Many of these observations correlate with the maturation of physiological response properties and suggest issues for further study.

Interaction of Excitation and Inhibition in Anteroventral Cochlear Nucleus Neurons That Receive Large Endbulb Synaptic Endings

Spherical bushy cells (SBCs) of the anteroventral cochlear nucleus (AVCN) receive their main excitatory input from auditory nerve fibers (ANFs) through large synapses, endbulbs of Held. These cells are also the target of inhibitory inputs whose function is not well understood. The present study examines the role of inhibition in the encoding of low-frequency sounds in the gerbil's AVCN. The presynaptic action potentials of endbulb terminals and postsynaptic action potentials of SBCs were monitored simultaneously in extracellular single-unit recordings in vivo. An input-output analysis of presynaptic and postsynaptic activity was performed for both spontaneous and acoustically driven activity. Two-tone stimulation and neuropharmacological experiments allowed the effects of neuronal inhibition and cochlear suppression on SBC activity to be distinguished. Ninety-one percent of SBCs showed significant neuronal inhibition. Inhibitory sidebands enclosed the high- or low-frequency, or both, sides of the excitatory areas of these units; this was reflected as a presynaptic to postsynaptic increase in frequency selectivity of up to one octave. Inhibition also affected the level-dependent responses at the characteristic frequency. Although in all units the presynaptic recordings showed monotonic rate-level functions, this was the case in only half of the postsynaptic recordings. In the other half of SBCs, postsynaptic inhibitory areas overlapped the excitatory areas, resulting in nonmonotonic rate-level functions. The results demonstrate that the sound-evoked spike activity of SBCs reflects the integration of acoustically driven excitatory and inhibitory input. The inhibition specifically affects the processing of the spectral, temporal, and intensity cues of acoustic signals.

The effects of congenital deafness on auditory nerve synapses: Type I and Type II multipolar cells in the anteroventral cochlear nucleus of cats.

Sensory deprivation has been shown to exert detrimental effects on the structure and function of central sensory systems. Congenital deafness represents an extreme form of auditory deprivation, and in the adult white cat, synapses between auditory nerve endings and resident cells of the anteroventral cochlear nucleus exhibited abnormal structure. Endbulbs of Held were reduced in branching and displayed striking hypertrophy of their postsynaptic densities. So-called modified endbulbs showed no change in branching complexity but did exhibit hypertrophy of their postsynaptic densities. These differential pre- and postsynaptic effects prompted the question of how deafness might affect other primary endings and synapses. Thus, we studied type I and type II multipolar cells that receive bouton endings from auditory nerve fibers. Type I multipolar cells project to the contralateral inferior colliculus and have relatively few axosomatic endings; type II multipolar cells project to the contralateral cochlear nucleus and have many axosomatic endings. Compared with normal-hearing cats, bouton endings of congenitally deaf cats were smaller but there was no difference in synaptic vesicle density or size of postsynaptic densities. These data reveal that different classes of primary endings and second-order neurons exhibit different degrees of synaptic anomalies to deafness.

Ultrastructural basis of synaptic transmission between endbulbs of Held and bushy cells in the rat cochlear nucleus.

Auditory nerve fibres make large excitatory synaptic contacts, the endbulbs of Held, with bushy cells in the anteroventral cochlear nucleus (AVCN). We have used serial-section electron microscopy to reconstruct seven endbulbs of Held in contact with three different AVCN bushy cells from a 25-day-old rat, as a basis for interpreting our previous physiological results at this connection. Four endbulbs of Held contacting the same bushy cell were completely reconstructed. The number of separate synaptic specializations within these endbulbs varied from 85 to 217, with a mean of 155. Detailed measurements were obtained from high magnification segments of four endbulbs contacting three different bushy cells. Large variability was found in the size of synaptic specializations within an individual endbulb. The size of postsynaptic densities (PSDs) varied between endbulbs (mean PSD area 0.03, 0.07, 0.07 and 0.18 microm(2); n = 4 endbulbs). The number of morphologically docked vesicles at individual specializations within the same endbulb varied considerably (between 1 and 102). The mean number of morphologically docked vesicles per specialization differed between endbulbs (mean numbers of docked vesicles per specialization = 2.1, 3.7, 5.3, 14.8; n = 4 endbulbs). Despite these large differences, the density of docked vesicles per square micron of PSD was similar between endbulbs (54, 80, 81, 83 docked vesicles per microm(2); n = 4 endbulbs). Within an endbulb, a linear relationship was found between the number of docked vesicles and PSD area, and between PSD area and the number of undocked vesicles within 150 nm of the active zone. The ratio of undocked vesicles (< 150 nm) to docked vesicles ranged from 2 to 5 in different endbulbs (n = 4 endbulbs). These structural observations are discussed in relation to the functional properties of synaptic transmission between endbulbs of Held and bushy cells in the AVCN.

The effects of congenital deafness on auditory nerve synapses and globular bushy cells in cats

It is well known that auditory deprivation affects the structure and function of the central nervous system. Congenital deafness represents one form of deprivation, and in the adult white cat, it has been shown to have a clear effect upon the synaptic interface between endbulbs of Held and spherical bushy cells. It is not known, however, whether all primary synapses are affected and/or whether they are affected in the same way and to the same extent. Thus, we studied a second neuronal circuit in the deaf white cat involving modified (small) endbulbs and globular bushy cells. Compared to normal hearing cats, modified endbulbs of congenitally deaf cats were 52.2% smaller but unchanged in structural complexity. There was also a striking loss of extracellular space between ending and cell body. The somata of postsynaptic globular bushy cells were 13.4% smaller and had enlarged postsynaptic densities. These data reveal that axosomatic synapses demonstrate abnormal structure as a consequence of deafness and that the extent of the abnormalities can vary with respect to the circuits involved. The implication of these observations is that synaptic anomalies would introduce differential delays within separate circuits, thereby desynchronizing neural activity from sound stimuli. This loss of synchronization could in turn disrupt temporal processing and compromise a host of related functions, including language comprehension.

Phosphorylation regulates spontaneous and evoked transmitter release at a giant terminal in the rat auditory brainstem.

1. The role of phosphorylation in synaptic transmission was investigated at a large glutamatergic terminal, the endbulb of Held, on bushy cells in the rat anteroventral cochlear nucleus (AVCN). 2. Whole-cell recordings of excitatory postsynaptic currents (EPSCs) were used to examine the effects of kinase inhibitors and activators on low-frequency (baseline) evoked release, spontaneous release, paired-pulse facilitation (PPF) or depression (PPD), repetitive stimuli and recovery from depression. 3. Application of the kinase inhibitor H7 (100 microM) reduced low-frequency evoked EPSC amplitude (by 15 %) and simultaneously increased PPF (or reduced PPD), with no significant change in other aspects of transmission. H7 did not affect the amplitude or frequency of spontaneous miniature EPSCs. 4. Phorbol esters increased EPSC amplitude (by 50 %) with a concomitant decrease in PPF (or increase in PPD), and reduced the final EPSC amplitude during repetitive stimuli. The effect of phorbol esters was due exclusively to protein kinase C (PKC) activation, as the specific PKC inhibitor bis-indolylmaleimide (Bis) completely blocked the potentiating effect of phorbol esters on EPSC amplitude. 5. Significantly, phorbol esters did not increase the evoked EPSC amplitude at connections in which release was maximized using high extracellular calcium concentrations (4-6 mM). 6. Phorbol esters increased the frequency of spontaneous miniature EPSCs in physiological calcium (by 275 %), and in high extracellular calcium (by 210 %) when phorbol esters did not increase the evoked EPSC amplitude. 7. Our results are most consistent with the actions of H7 to decrease low-frequency release probability and phorbol esters to increase low-frequency release probability at the endbulb-bushy cell synaptic connection in the AVCN. The effects of H7 and phorbol esters on paired-pulse responses and tetanic depression appear to be largely consequential to these changes in low-frequency release probability.

A novel presynaptic inhibitory mechanism underlies paired pulse depression at a fast central synapse.

Several distinct mechanisms may cause synaptic depression, a common form of short-term synaptic plasticity. These include postsynaptic receptor desensitization, presynaptic depletion of releasable vesicles, or other presynaptic mechanisms depressing vesicle release. At the endbulb of Held, a fast central calyceal synapse in the auditory pathway, cyclothiazide (CTZ) abolished marked paired pulse depression (PPD) by acting presynaptically to enhance transmitter release, rather than by blocking postsynaptic receptor desensitization. PPD and its response to CTZ were not altered by prior depletion of the releasable vesicle pool but were blocked by lowering external calcium concentration, while raising external calcium enhanced PPD. We conclude that a major component of PPD at the endbulb is due to a novel, transient depression of release, which is dependent on the level of presynaptic calcium entry and is CTZ sensitive.

Development of dynorphin-like immunoreactive auditory nerve terminals in the chick

The novel discovery that auditory nerve terminals in the chick cochlear nucleus magnocellularis (NM) are immunoreactive for the opioid peptide dynorphin (DYN) was recently reported [3]. The present study examines the development of DYN-immunoreactivity (DYN-I) in auditory nerve terminals in NM from embryos through young post-hatch chicks. No DYN-I was observed in NM at embryonic day 13 (E13). DYN-I first appeared at E16 as short flat structures partially surrounding NM cell bodies. Around post-hatch day 1 (P1), these structures had a more rounded, chalice-type of morphology reminiscent of the specialized auditory nerve terminals found in birds, the end-bulbs of Held. At P6, most NM neurons were circumscribed by a prominent DYN-I calyceal-type of ending. By P13, fewer NM cells were ringed by this DYN-I and by the third post-hatch week, there was very little DYN-I in NM. There were no obvious differences in the density of DYN-I terminals across either the rostrocaudal length or the mediolateral width of NM at any age examined. These results suggest that during a restricted time of development, end-bulbs of Held in the chick NM contain DYN.

Chick auditory terminals contain dynorphin-like immunoreactivity.

Dynorphin-like immunoreactivity (DYN-I) was studied in the auditory brain stem of chick hatchlings using an antiserum to dynorphin-B. DYN-I was found in a few small cells in the nucleus laminaris (NL) and in the superior olive (SO). Dendrites in NL also appeared immunostained. DYN-I, however, was especially prominent in the cochlear nucleus magnocellularis (NM). A few DYN-I somata could be found within NM particularly in medial regions of this nucleus. Most DYN-I, however, was localized to calyceal-type structures, reminiscent of primary auditory nerve terminals, surrounding nerve cell bodies in NM. To determine whether DYN-I is contained within auditory end-bulbs of Held, the basilar papilla (avian cochlea) was removed unilaterally. After 5 days survival, there were very few DYN-I terminals in NM on the operated side of the brain compared with the unoperated side. This novel finding suggests that primary auditory nerve terminals in NM may contain the opioid peptide, dynorphin.

Activity-related features of synapse morphology: a study of endbulbs of held.

The myelinated fibers of the auditory nerve can be divided into two separate populations on the basis of sensitivity to sound, average levels of spike activity, and central branching patterns. The synaptic endings of these populations were investigated for the presence of structural specializations that might correlate with levels of neural activity. We applied intracellular recording and staining methods in cats to analyze directly the relationship between spike activity and the structure of synapses using endbulbs of Held, the large synaptic endings in the anteroventral cochlear nucleus. Endbulbs from fibers having low or high levels of activity were examined and compared using light and electron microscopic methods. All endbulbs exhibited relatively large but incomplete coverage by one-to-several lamellae of glial processes. Endbulbs of high activity fibers were large and contained larger mitochondria than endbulbs of low activity fibers. Furthermore, the synapses of high activity endbulbs were on average smaller but more numerous, possessed greater numbers of associated synaptic vesicles, and exhibited greater curvature of their postsynaptic densities. These structural features are hypothesized to reflect specializations that optimize synaptic transmission.

Receptors underlying excitatory synaptic transmission in slices of the rat anteroventral cochlear nucleus

1. The anteroventral cochlear nucleus (AVCN) contains two principal cell types that receive input from the auditory nerve. Stellate cells receive conventional synapses on their dendrites, and bushy cells of the AVCN receive axosomatic input via large, calyceal terminals (the end bulbs of Held). We have used whole cell patch-clamp recording techniques to study excitatory postsynaptic currents (EPSCs) in these two principal cells of the rat AVCN. 2. EPSCs evoked in stellate cells by stimulation of the auditory nerve were graded with stimulus strength, indicating a high degree of convergence of input to these cells. At depolarized membrane potentials, EPSCs evoked in stellate neurons had a dual-component time course. The slow component was blocked by the N-methyl-D-aspartate (NMDA) receptor antagonist DL-2-amino-5-phosphonovaleric acid (APV), and the fast component was abolished by the non-NMDA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX). 3. EPSCs evoked in bushy cells by auditory nerve stimulation were large (50 nS average conductance) and all-or-none at the threshold stimulus level. At -70 mV, the time course of the EPSC was very brief (average time constant of decay 700 microseconds at room temperature). Membrane depolarization revealed a slow component to the EPSC. The fast and slow components were mediated by non-NMDA and NMDA receptors, respectively. The switch-off of end bulb NMDA EPSCs by voltage jumps to the EPSC reversal potential was very rapid, suggesting that the NMDA component arises from sites on or close to the soma. 4. Miniature EPSCs, recorded in the presence of tetrodotoxin (TTX) at depolarized potentials, also had a dual-component time course. The fast and slow components of the miniature EPSCs were blocked by CNQX and APV, respectively. This result indicates that NMDA and non-NMDA receptors can be co-localized at the same, presumably end bulb, release sites. 5. The relative contribution of the slow, NMDA component to the end bulb EPSC declined significantly with age (postnatal days 11-22). 6. These results indicate that both NMDA and non-NMDA receptors underlie excitatory synaptic transmission in the AVCN of young rats. The end bulb synapse onto bushy cells generates a non-NMDA receptor-mediated EPSC with very fast kinetics. NMDA receptors can also mediate synaptic transmission at the end bulb synapse, but their contribution becomes less as the auditory system matures. This finding suggests that NMDA receptors may play an important role in the development of this synapse.

Anatomical evidence of synaptic plasticity in the cochlear nuclei of white-deaf cats

Synapses are dynamic structures reflecting environmental events. It was observed that densities associated with apposing synaptic membranes were altered in response to changing conditions of auditory stimulation ( Gulley et al. 1978 . J. Comp. Neurol. 180: 707–742). Synapses also exhibit plasticity during early development of the auditory system ( Larsen and Pappas, 1985. Proceedings: 43rd Annual Meeting of the Electron Microscopy Society of America, pp. 493–494). Larsen and Pappas reported that, in young kittens, the end bulb of Held (EBH) terminal was distinguished by excessive density at apposing synaptic membranes which assume gradually the pattern observed in adult cats. The maturation of synapses parallels emerging function and may depend upon auditory stimulation ( Larsen and Kirchhoff, 1987. Neurosci. Abstr. 13: 1260). Larsen and Kirchhoff found that EBH synapses on large spherical cells of white-deaf cats resembled the immature synapses found in young kittens (Fig. 1). They also found terminals that had an increased number and length of the membrane densities associated with synapses. Because the number of synaptic vesicles in these presynaptic terminals was equivalent to the number of vesicles found in normal-hearing cats, Larsen and Kirchhoff (1987) suggest that this is evidence of synaptic plasticity. We have completed a quantitative study of the synapses of EBH terminals found in adult cats that had been deaf for at least 3 years.

Central projections of auditory nerve fibers in the barn owl

The central projections of the auditory nerve were examined in the barn owl. Each auditory nerve fiber enters the brain and divides to terminate in both the cochlear nucleus angularis and the cochlear nucleus magnocellularis. This division parallels a functional division into intensity and time coding in the auditory system. The lateral branch of the auditory nerve innervates the nucleus angularis and gives rise to a major and a minor terminal field. The terminals range in size and shape from small boutons to large irregular boutons with thorn-like appendages. The medial branch of the auditory nerve conveys phase information to the cells of the nucleus magnocellularis via large axosomatic endings or end bulbs of Held. Each medial branch divides to form 3-6 end bulbs along the rostrocaudal orientation of a single tonotopic band, and each magnocellular neuron receives 1-4 end bulbs. The end bulb envelops the postsynaptic cell body and forms large numbers of synapses. The auditory nerve profiles contain round clear vesicles and form punctate asymmetric synapses on both somatic spines and the cell body.

Synaptic connections of the auditory nerve in cats: Relationship between endbulbs of held and spherical bushy cells

This report focuses on a class of large synaptic endings, the endbulbs of Held. These endings are located in the anteroventral cochlear nucleus and arise from the axons of type I spiral ganglion neurons. Axons were stained with horseradish peroxidase (HRP) using intracellular injections of single fibers or extracellular injections into the auditory nerve. Individual endbulbs or pairs of endbulbs that converged onto the same spherical bushy cell were examined with the aid of a light microscope and subjected to morphometric analyses. Endbulbs of fibers having low spontaneous discharge rates (SR, less than or equal to 18 spikes/sec) have a more complex shape than those of high SR fibers (greater than 18 s/s), a feature represented by systematic differences in endbulb silhouette perimeter without differences in endbulb silhouette area. Consequently, the ratio, silhouette area divided by silhouette perimeter, yields a "form factor" separating endbulbs of high SR from those of low SR. High SR fibers had ratios greater than 0.52 (mean = 0.63 +/- 0.09), whereas low SR fibers had ratios less than 0.52 (mean = 0.45 +/- 0.06). Pairs of endbulbs with unknown physiological properties had similar form factor values, despite the wide range of values observed in the endbulb population. These data imply that endbulbs converging upon the cell body of a spherical bushy cell arise from fibers of the same SR group. Electron microscopic examination was conducted on the endbulb of one physiologically characterized and intracellularly stained auditory nerve fiber (CF = 1.4 kHz; SR = 55 s/s) and its unstained endbulb mate with the aid of serial ultrathin sections. In addition to the well-known axosomatic synapses, these endbulbs formed axodendritic synapses: 11.7% for the HRP-labeled endbulb and 13.3% for the unlabeled endbulb. The axodendritic synapses appear to occur on dendrites of nearby spherical bushy cells and may represent a mechanism whereby single endbulbs can disperse activity to multiple neurons in the cochlear nucleus. We propose that axosomatic synapses preserve fiber SR groupings, whereas axodendritic synapses may not.

The nucleus magnocellularis in the red-eared turtle, Chrysemys scripta elegans: Eighth nerve endings and neuronal types

Eighth nerve endings and neuronal types in the nucleus magnocellularis were analyzed in the red-eared turtle ( Chrysemys scripta elegans). One group of turtles had horseradish peroxidase (HRP) injected into the surgically exposed inner ear. Following injection the animals survived for 3–5 days and then their 8th nerve fibers and endings were analyzed. A second group of turtles were impregnated by the Golgi-Kopsch technique. This also demonstrated 8th nerve endings and neurons in the nucleus magnocellularis. A third group had brains stained with cresyl violet to demonstrate normal morphology. Two types of neurons were present in the nucleus magnocellularis; bushy and stellate. Bushy neurons had a single primary dendrite with little branching and spines. Stellate neurons had 3–5 primary dendrites with secondary order branching and dendritic spines. Thick 8th nerve axons gave rise to endbulbs of Held and their axons formed bouton-type ending. Endbulbs of Held were presently only on bushy neurons while bouton-type terminals were only on stellate neurons. Endbulbs of Held had two patterns; one group was cup-shaped and surrounded approximately 1 2 of the soma without appendages. A second type had a smaller cup with 3–5 extensions to other parts of the cell. Stellate neurons had terminal boutons adjacent to the cell membrane. They appeared in some situations as a necklace surrounding the neuron.

Central projections of intracellularly labeled auditory nerve fibers in cats: morphometric correlations with physiological properties.

The central arborizations and endings of type I spiral ganglion neurons were labeled with intracellular injections of horseradish peroxidase (HRP) after their characteristic frequency (CF) and spontaneous discharge rate (SR) were physiologically determined. A fiber-by-fiber analysis was conducted and the morphological data compared with the fiber's response properties. The total number of branch points was correlated with total fiber length, a relationship that remained relatively constant when analyzing the ascending and descending branches together or separately. On the other hand, the ascending branches of four out of five fibers having CFs below 0.5 kHz bifurcated and gave rise to a pair of terminal endbulbs of Held. Low- and medium-SR fibers gave rise to more endings than did high-SR fibers, especially on the ascending branch. This difference was accounted for by small endings, a category composed of terminal boutons, string endings, and small complex endings. The categories of modified endbulbs, and endbulbs of Held did not vary in number with respect to fiber SR. The mean area of each ending type within the small ending category was statistically smaller for low- and medium-SR fibers than for high-SR fibers, whereas the mean area of modified endbulbs and endbulbs of Held was not correlated with fiber SR. Total ending area per fiber appeared independent of either CF or SR. These results are discussed in relation to issues of conservation of axon arborizations and terminals, and convergence of input from the different SR groups.

The central projections of intracellularly labeled auditory nerve fibers in cats: an analysis of terminal morphology.

The axons of physiologically characterized spiral ganglion neurons (type I) were stained throughout their arborizations in the cochlear nucleus by the intracellular injection of horseradish peroxidase (HRP). The tips of the axonal branches were marked by distinct swellings, ranging in size and shape from small boutons to large perisomatic ramifications. Electron microscopic analysis of such swellings revealed ultrastructural features characteristic of primary auditory synapses, consistent with the hypothesis that terminal swellings identifiable in the light microscope represent presynaptic endings. On the basis of light microscopic differences in size, these endings were organized into three categories. Endings of relatively small size (terminal boutons, free endings, boutons with filopodia, string endings, and small complex endings) composed 94% of all terminal endings. Within this category of small endings, there were predictable variations in relative size and regional distribution that related to the spontaneous discharge rate (SR) of the fiber. The endings of low and medium SR fibers (SR less than or equal to 18 spikes/second) were smaller on average than those of high SR fibers (SR greater than 18 spikes/second). Furthermore, there were more endings arising from the ascending branch than from the descending branch when comparing fibers of the low and medium SR group with those of the high SR group. There were not, however, obvious morphological features of this ending category that correlated with the characteristic frequency (CF, the pure tone frequency to which the neuron is most sensitive). A second category contained medium-sized complex endings, most of which formed axosomatic contacts. This category composed 4% of the population and was found in close proximity to the perikarya of globular, octopus, and spherical cells. The endings from low and medium SR fibers were smaller on average than those from high SR fibers. These endings did not vary in their parent branch distribution with respect to fiber SR, nor did they exhibit morphological features that correlated with fiber CF. The third category contained large complex endings (endbulbs of Held) and composed 2% of the ending population. Within the anteroventral cochlear nucleus, these large, complex endings made axosomatic contact with spherical cells in the anterior division and with globular cells in the posterior division. There were no systematic variations in ending size or branch distribution that correlated with fiber SR. There was, however, a relationship between ending size and fiber CF such that fibers having CFs below 4 kHz gave rise to the largest endbulbs.